CN1328260A - Integrated immune biochip, electric signal pick-up interface and its preparing process - Google Patents
Integrated immune biochip, electric signal pick-up interface and its preparing process Download PDFInfo
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- CN1328260A CN1328260A CN 01118727 CN01118727A CN1328260A CN 1328260 A CN1328260 A CN 1328260A CN 01118727 CN01118727 CN 01118727 CN 01118727 A CN01118727 A CN 01118727A CN 1328260 A CN1328260 A CN 1328260A
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Abstract
An integrated biologic immune chip is prepared through making the metal films on two substrates become multiple electrodes with a certain shape, area and interval, the leading lines for connecting said electrodes, and the terminals connected with said leading lines, adsorbing the bioimmune antibody on the electrodes, correctly positioning said electrodes by face to face, and packing it in a U-shaped container. Its electric signal pick-up interface is prepared from flexible circuit board and rubber board. Its advantages are high test speed and accuracy.
Description
The invention provides a kind of integrated immune biochip, electric signal pick-up interface and manufacturing technology thereof.A kind of method that has adopted even glue, photoetching, etching technics in coating technique and the SIC (semiconductor integrated circuit) manufacturing technology to make this integrated immune biochip particularly is provided; A kind of method that adopts flex circuit application and flexible rubber plate to be combined into special-purpose electric signal pick-up interface is provided.
Modern medicine is told us, and many malignant tumours all have a transition process from optimum to pernicious.Because existing many medical detection means sensitivity are lower, can not just make correct diagnosis at the tumour initial stage that changes, thereby miss the most effective treatment opportunity, incured loss through delay patient's life.And the biological immune chip utilizes the immunology principle can be from the incunabulum of patient morbidity, or even judge from formation stage of disease immune state to human body, thereby the various diseases of human body especially resembles some difficult and complicated illness such as cancer, angiocardiopathy and diabetes.The principle of biological immune chip detection is to utilize the antigen of certain disease and the uniqueness that combines with its corresponding antibodies, that is the cohesive process of different types of antigen (virus, bacterium, toxin etc.) and its corresponding antibodies has specificity and exclusiveness, analyze in the human body quantity with certain disease corresponding antigen, thereby the possibility that human body will produce certain disease is judged.A canonical process of biological immune chip preparation at present is: at first go up the membraneous material (material commonly used at present is a gold) that plating one deck can adsorb certain activator at certain substrate (as: glass or silicon chip), make it to adsorb one deck biomolecule activator through handling, the antibody of the various diseases that will detect is adsorbed onto on the active layer with chemical method then, just constitutes the biological immune chip.Generally be used for when clinical when the biological immune chip, suppose to be adsorbed on equably on the active layer of chip with certain disease corresponding antibody, whether the blood sample with tested human body is coated on the chip then, have, have the antibody of certain disease on how much antigen and the chip to combine again on the detection chip.The correctness that homogeneity that antibody distributes on the chip and compactness directly have influence on diagnostic result.Have only this measurement accurate, could make correct judgement to the health status of human body, therefore, the key of biological immune chip technology is: 1. improve the density that antibody distributes on the chip; 2. improve the homogeneity that antibody distributes on the chip, 3. accurately read antigen number with antibodies.Widely used in the world at present biological immune chip detection technology mainly contains methods such as atomic force microscope (AFM), chemiluminescence, beautiful fluorescence and plasma resonance.Several method detection mode difference respectively has its advantage, but all has certain limitation.For example utilize AFM, can observe the pattern of biomolecule intuitively, but shortcoming is that not only instrument itself costs an arm and a leg, and detection speed is very limited.Chemiluminescence method because the fluorescent lifetime of used luminescent material is limited, is difficult to detect accurately.Beautiful fluorescent method be at present in the biological immune chip detection relatively effectively and the method that is widely used, it is based on antagonist and carries out certain fluorized marking, thereby realizes detection to antigen quantity by detecting fluorescence signal intensity.Its advantage is that principle is simple, and detection means is more direct.But its maximum shortcoming is to detect very difficulty when not only realizing multiple disease, and because antagonist carries out fluorized marking process complexity, cost is very high.The application of plasma resonance method also is subjected to the restriction of detection speed and instrument cost.
Order of the present invention is to provide a kind of integrated immune biochip, electric signal pick-up interface and preparation method thereof.Realize quick, accurately detection.
Formation of the present invention:
1, integrated immune biochip is made up of substrate and the metallic film that generates on substrate.Substrate can adopt amorphous material or macromolecular material, can also adopt metal oxide ceramic material or semiconductor material.Metallic film can be the elemental metals film; It also can be the alloying metal film; It also can be metal-oxide film; It can also be metallized multilayer film.The thickness of metallic film is 20 nanometers---5000 nanometers.With the even glue in the SIC (semiconductor integrated circuit) manufacturing technology, photoetching, etching technics metallic film make electrode, contact conductor that is connected with each electrode and the electrode external terminal that is connected with each contact conductor.Make two on-chip electrode absorption go up biological immune antibody., employing mark line location face-to-face one by one each electrode that is adsorbed with biological immune antibody on two substrates placed and is encapsulated in the vessel that a three side sealing closes, the U font wall of recording quantity constitutes, and the external measurement terminal of electrode is positioned at the outside of vessel.Thereby make integrated immune biochip.
2, electric signal pick-up interface is made up of flex circuit application and flexible rubber plate.One end of flex circuit application is made the complete corresponding solder joint circuitous pattern of feature size, quantity and spacing with the electrode external terminal of integrated immune biochip of the present invention.Solder joint is the Gold plated Layer that size is even, thickness equates, plated thickness is 0.02 millimeter---2 millimeters.The other end of flex circuit application is widened the size that is connected mutually with universal multipole plug wire terminal by lead-in wire.At the back side of gold-plated solder joint one side of flex circuit application liner lastblock flexible rubber plate.When flex circuit application and flexible rubber plate are evenly exerted pressure, the gold-plated solder joint of each on the flex circuit application will keep the excellent contact conducting with the external measurement terminal of its corresponding electrode.Thereby make electric signal pick-up interface.
3, make the method for above-mentioned integrated immune biochip by selecting substrate, adsorb biological immune antibody in the shaping of the electrode external terminal for preparing metallic film, electrode and the contact conductor that is connected with each electrode on the substrate and be connected, on metal electrode, making a vessel that includes the electrode that all is adsorbed with biological immune antibody with each contact conductor.Specific embodiment is for selecting substrate → preparation metallic film → shaping → absorption biological immune antibody → making vessel.
(1) select substrate: substrate can be amorphous materials such as biological slide, simple glass, silicon dioxide; Also can be plastics, macromolecular materials such as poly-four compounds; Also can be zinc paste, barium titanate, lead titanates, lead zirconate titanate metal oxide ceramic materials such as (PZT); Can also be semiconductor materials such as silicon, gallium arsenide.
(2) prepare metallic film: the method with physical deposition or chemogenic deposit on substrate plates 20 nanometers---the metallic film of 5000 nanometer thickness.Can adopt vacuum deposition method, direct current/radio frequency plasma magnetron sputtering method, electron beam vacuum deposition method, laser to melt the method for physical deposition method such as method, ion beam sputtering, molecular beam epitaxial growth method or metal-organic chemical vapor deposition equipment method (MOCVD), sol-gel process (Sol-Gel), metal-organic decomposition method chemogenic deposits such as (MOD), metal-coated films on above-mentioned substrate.The metallic film that is deposited can be Au, Al, Pt, Ir, Cu, Ag, Ni, Co, elemental metals films such as Fe; Also can be AuCu, NiCo, NiFe, PtCu, alloying metal films such as CuAg; Also can be iridium dioxide (IrO2), ruthenic oxide metal-oxide films such as (RuO2); Can also be metallized multilayer films such as Au/Cr, Au/Al, Au/Ti.
(3) forming technology: after adopting above-mentioned coating technique to make various metallic films, with the even glue in the SIC (semiconductor integrated circuit) manufacturing technology, photoetching, etching technics metallic film make electrode, contact conductor that is connected with each electrode and the electrode external terminal that is connected with each contact conductor.The etching manufacturing process, as: ion etching method, chemical erosion method, laser ablation method, mould forming method, mask method etc.
(4) absorption biological immune antibody: utilize the bioactivation technology that the electrode that adopts above-mentioned moulding process and make is carried out surface activation process, absorption one deck activating substance on electrode, the biological immune antibody of the various diseases that will detect is adsorbed onto on the active layer with chemical method then.
(5) make vessel:, employing mark line location face-to-face one by one each electrode that is adsorbed with biological immune antibody on two substrates placed and is encapsulated in the vessel that a three side sealing closes, the U font wall of recording quantity constitutes, and the external measurement terminal of electrode is positioned at the outside of vessel, thereby makes integrated immune biochip.The encapsulating material of making the U font wall of vessel can be an epoxy resin, also can be plastics, poly-four compounds and elastomeric material etc.Distance between two substrates is from 0.05 millimeter---and 10 millimeters.
4, the method for the aforementioned electric signal pick-up interface of making is as follows:
One end of flex circuit application is made the complete corresponding solder joint circuitous pattern of feature size, quantity and spacing with the external measurement terminal of electrode of integrated immune biochip of the present invention.Solder joint is the Gold plated Layer that size is even, thickness equates, plated thickness is 0.05 millimeter---2 millimeters.The other end of flex circuit application is widened the size that is connected mutually with universal multipole plug wire terminal by lead-in wire.At the back side of gold-plated solder joint one side of flex circuit application liner lastblock flexible rubber plate.When flex circuit application and flexible rubber plate are evenly exerted pressure, the gold-plated solder joint of each on the flex circuit application will keep the excellent contact conducting with the external measurement terminal of its corresponding electrode.Thereby make electric signal pick-up interface.In the above-mentioned electric signal pick-up interface, the solder joint coating material on the flex circuit application can also adopt materials such as silver, copper, aluminium, nickel, scolding tin except adopting gold.In the perhaps above-mentioned electric signal pick-up interface, the gold-plated solder joint of each on the flex circuit application also can use the pressure line terminal with the built-in miniature spring of the external measurement terminal equal number of electrode to replace.
The invention has the advantages that: detection method is compared with the detection method of the biological immune chip of present use, it is more rapid to have detection speed, testing result is more accurate, the checkout equipment volume reduces, in light weight, advantage such as cost significantly reduces than aforementioned other method, and disposable use consumptive material is cheap, be easy to also realize that multiple disease once detects simultaneously, and can realize large batch of clinical medical detection.
Below in conjunction with accompanying drawing furtherly to the present invention:
Fig. 1 is the section stratiform structural representation that the present invention makes the metallic gold film that integrated immune biochip uses.Substrate is a biological slide.(1) gold thin film, (2) biological slide.
Fig. 2 is the photoresist section stratiform structural representation of the electrode shape that forms on the metallic gold film of the present invention.Substrate is a biological slide.(1) gold thin film, (2) biological slide, (3) photoresist.
Fig. 3 is the section stratiform structural representation of metallic gold membrane electrode among the present invention.Substrate is a biological slide.(2) biological slide, (4) gold film electrode.
Fig. 4 is the floor map of metallic gold membrane electrode of the present invention, gold film electrode lead-in wire and the external measurement terminal of gold film electrode.Substrate is a biological slide.(2) biological slide, (4) gold film electrode, (5) gold film electrode lead-in wire, the external measurement terminal of (6) gold film electrode.
Fig. 5 is the section stratiform structural representation that is adsorbed with biological immune antibody among the present invention on the metallic gold membrane electrode.Substrate is a biological slide.(2) biological slide, (4) gold film electrode, (7) activating substance layer, (8) biological immune antibody.
Fig. 6 is the structural representation of integrated immune biochip of the present invention (10).(2,2 ') biological slide, (4,4 ') gold film electrode, (5,5 ') gold film electrode lead-in wire, (6,6 ') gold film electrode external terminal, (9) U font wall.
Fig. 7 is the synoptic diagram of the electric signal pick-up interface (17) be made up of flex circuit application and flexible rubber plate among the present invention.(11) flex circuit application, (12) gold-plated solder joint, (13) widen contact conductor, (14) flexible rubber plate, (15) universal multipole plug wire terminal, (16) plug wire post.
Fig. 8 is the section structure synoptic diagram after integrated immune biochip of the present invention (10) and electric signal pick-up interface (17) assembling.(2,2 ') biological slide, (6,6 ') gold film electrode external terminal, (9) U font wall, (11,11 ') flex circuit application, (12,12 ') gold-plated solder joint, (14,14 ') flexible rubber plate, the aggregate of (18,18 ') gold film electrode, activating substance layer and biological immune antibody.Upper and lower two thick arrows represent to apply the direction of uniform pressure.
Fig. 9 is the assembling synoptic diagram after the pressure line terminal of built-in miniature spring among the present invention replaces flex circuit application.(2,2 ') biological slide, (4,4 ') gold film electrode, (5,5 ') gold film electrode lead-in wire, (6,6 ') gold film electrode external terminal, (9) U font wall, the pressure line terminal of (19) built-in miniature spring, (20) external wire.
Figure 10 is the synoptic diagram of the pressure line terminal of built-in miniature spring of the present invention.(20) external wire, (21) shell, (22) built-in miniature spring, (23) movable pressure line terminal.
Go up the biological integrated chip of making in biological slide (glass substrate).
To long 45 millimeters, wide 22 millimeters, thick 0.8 millimeter biological slide with the acetone ultrasonic cleaning after, use the deionized water ultrasonic cleaning, use the absolute ethyl alcohol ultrasonic cleaning at last.Biological slide after the cleaning dries up with high pure nitrogen.
The biological slide of cleaning is put in the coating chamber, coating chamber is extracted into 2 * 10
-4The vacuum tightness of Pa.On biological slide, deposit the gold thin film of 200 nanometer thickness with vacuum coating equipment.Vacuum tightness 3 * 10 during deposition
-3Pa, substrate temperature 200 degree.Gold thin film thickness can be controlled with the adjustment sedimentation time.The section layer structure of metallic gold film as shown in Figure 1.
After the gold thin film deposition is finished, photoresist is coated in the gold thin film surface.Use has the photolithography plate of electrode, contact conductor and electrode external terminal shape, that is, electrode shape is 0.4 millimeter circle of diameter, and lead format is the elongate strip of 0.02 millimeter of width, and the electrode external terminal is that length and width are respectively 0.4 millimeter square.Photolithography plate covered as shadow shield scribbling on the photoresist gold thin film, make the photoresist photosensitive hardening that is not covered by shadow shield with ultraviolet ray, then, with the chemical solution flush away not by the photoresist of UV-radiation-sensitive.So, elongate strip of 0.02 millimeter of 0.4 millimeter circle of diameter, width and the foursquare photoresist overlay area that length and width are respectively 0.4 millimeter have been formed on the gold thin film surface, referring to Fig. 2.Fig. 2 has only provided the photoresist overlay area at electrode place.After this sample carried out etching with ion etching equipment, the elongate strip gold film electrode lead-in wire and the length and width that have just formed 0.02 millimeter of 0.4 millimeter circular gold film electrode of diameter and width are respectively 0.4 millimeter foursquare gold film electrode external terminal, referring to Fig. 3 and Fig. 4.
Utilize the bioactivation technology that the gold film electrode that adopts above-mentioned moulding process and make is carried out activation processing, absorption one deck activating substance on electrode, the biological immune antibody of the various diseases that will detect is adsorbed onto on the active layer with chemical method then, referring to Fig. 5.
, employing mark line location face-to-face one by one each electrode that is adsorbed with biological immune antibody on two substrates placed and is encapsulated in the vessel that a three side sealing closes, the U font wall of recording quantity constitutes, and the external measurement terminal of electrode is positioned at the outside of vessel.Thereby make integrated immune biochip.The encapsulating material of making the U font wall of vessel can be an epoxy resin, also can be plastics, poly-four compounds and elastomeric material etc.Distance between two substrates is from 0.05 millimeter---and 10 millimeters.Referring to Fig. 6.In Fig. 6, be adsorbed with biological immune antibody (not drawing among the figure) between two gold film electrodes up and down.
Shape, size and the number of gold film electrode (4,4 ') is not subjected to the restriction of present embodiment.Shape, size and the bar number of gold film electrode lead-in wire (5,5 ') is not subjected to the restriction of present embodiment.Shape, size and the number of the external measurement terminal of gold film electrode (6,6 ') is not subjected to the restriction of present embodiment.The external measurement terminal of two on-chip gold film electrodes (6,6 ') up and down shown among Fig. 6 also can be at the vessel homonymy of two substrates (2,2 ') and U font wall (9) formation.
According to above-mentioned steps, make integrated immune biochip provided by the invention.
Make electric signal pick-up interface.
Make corresponding gold-plated solder joint (12) with each electrode external terminal of the integrated immune biochip (10) of embodiment 1 at an end of flex circuit application (11).That is, each gold-plated solder joint is the right cylinder of 0.4 millimeter of diameter, 0.3 millimeter of height.The other end of flex circuit application is equipped with universal multipole plug wire terminal (15).Each gold-plated solder joint (12) is widened with universal multipole plug wire terminal (15) by its corresponding lead-in wire (13) and is connected mutually.Back side liner lastblock flexible rubber plate (14) in gold-plated solder joint one side of flex circuit application.Referring to Fig. 7.
The size of the gold-plated solder joint on the flex circuit application and number are not subjected to the restriction of present embodiment.The shape of flex circuit application is not subjected to the restriction of present embodiment.
Embodiment 3
With the electric signal pick-up interface (17) that is made among the embodiment 2 as shown in Figure 8 precision positioning be pressed on the external measurement terminal (6 of electrode of the integrated immune biochip (10) that is made among the embodiment 1,6 ') on, in actual measurement, two substrates (2 about the integrated immune biochip, 2 ') and each electrode external terminals of the vessel both sides (left and right sides) that constitute of U font wall (9) press simultaneously to put and connect the power on signal pick-up interface, just can the while, transient state detects owing to the different caused change in electric of biological antibody with antigen combination degree and quantity.
With the electric signal pick-up interface among the embodiment 3 (promptly, flex circuit application and flexible rubber plate) use the pressure line terminal (19) of built-in miniature spring to replace, the external measurement terminal of each electrode (6,6 ') all uses the pressure line terminal (19) of a corresponding built-in miniature spring to contact on the integrated immune biochip.As shown in Figure 9.For the sake of clarity, Fig. 9 has only shown the pressure line terminal of a built-in miniature spring.Figure 10 has shown the detailed structure of the pressure line terminal (19) of built-in miniature spring.Telescopic movable pressure line terminal (23) in the pressure line terminal of built-in miniature spring is made with metal material is gold-plated.2 millimeters of 0.3 millimeter of the diameter of movable pressure line terminal (23), length.2 millimeters of 0.2 millimeter of the diameter of the built-in miniature spring (22) that control activity pressure line terminal is flexible, length.6 millimeters of the diameters of the shell of the pressure line terminal of built-in miniature spring (21), 8 millimeters of length.External wire (20) is the copper conductor in 0.5 square millimeter of cross section.In actual measurement, two substrates (2 about the integrated immune biochip, 2 ') and the external measurement terminal (6 of each electrode of the vessel both sides (left and right sides) that constitute of U font wall (9), 6 ') press simultaneously and put the pressure line terminal (19) that the built-in miniature spring is gone up in contact, just can be simultaneously, transient state detects owing to the different caused change in electric of biological antibody with antigen combination degree and quantity.
Claims (5)
1, a kind of integrated immune biochip, it is characterized in that: go up the preparation metallic film at substrate (2,2 '), with the even glue in the SIC (semiconductor integrated circuit) manufacturing technology, photoetching, etching technics metallic film make electrode, the contact conductor that is connected with each electrode reaches the electrode external terminal that is connected with each contact conductor; Make the electrode absorption on the substrate (2,2 ') go up biological immune antibody; , employing mark line location face-to-face one by one each electrode that is adsorbed with biological immune antibody on the substrate (2,2 ') placed and is encapsulated in the vessel that a three side sealing closes, the U font wall of recording quantity constitutes, and the external measurement terminal of electrode is positioned at the outside of vessel; Thereby make integrated immune biochip.
2, according to the described integrated immune biochip of claim 1, it is characterized in that: substrate (2,2 ') can be biological slide, simple glass, silicon dioxide amorphous material, also can be plastics, poly-four compound macromolecular materials, or zinc paste, barium titanate, lead titanates, lead zirconate titanate (PZT) metal oxide ceramic material can also be silicon, gallium arsenide semiconductor material; Prepared metallic film can be Au, Al, Pt, Ir, Cu, Ag, Ni, Co, Fe elemental metals film, also can be AuCu, AuCr, NiCo, NiFe, PtCu, CuAg alloying metal film, or iridium dioxide, ruthenic oxide metal-oxide film, can also be Au/Cr, Au/Al, the Au/Ti metallized multilayer film; The thickness of metallic film is 20 nanometers---5000 nanometers.
3, a kind of electric signal pick-up interface to the described integrated immune biochip of claim 1, it is characterized in that: adopt flex circuit application and flexible rubber plate to combine, one end of flex circuit application is made the complete corresponding solder joint circuitous pattern of feature size, quantity and spacing with the electrode external terminal of above-mentioned integrated immune biochip, all solder joints are the Gold plated Layer that size is even, thickness equates, plated thickness is 0.05 millimeter---2 millimeters; The other end of flex circuit application is widened the size that is connected mutually with universal multipole plug wire terminal by lead-in wire, at the back side of gold-plated solder joint one side of flex circuit application liner lastblock flexible rubber plate, when flex circuit application and flexible rubber plate are evenly exerted pressure, the gold-plated solder joint of on the flex circuit application each will keep the excellent contact conducting with the external measurement terminal of the electrode in its corresponding integrated immune biochip, thereby can accurately detect electric signal.
4, according to the described electric signal pick-up interface of claim 3, it is characterized in that: the solder joint coating material on the flex circuit application can adopt silver, copper, aluminium, nickel, scolding tin conductive material; The gold-plated solder joint of on the flex circuit application each also can use the pressure line terminal with the built-in miniature spring of the external measurement terminal equal number of electrode to replace.
5, a kind of method for making of integrated immune biochip is characterized in that: this method is by selecting substrate, adsorb biological immune antibody in the shaping of the electrode external terminal for preparing metallic film, electrode and the contact conductor that is connected with each electrode on the substrate and be connected with each contact conductor, on metal electrode, making a vessel that includes the electrode that all is adsorbed with biological immune antibody; Technological process is for selecting substrate → preparation metallic film → shaping → absorption biological immune antibody → making vessel.
According to the described method for making of claim 5, it is characterized in that:
A selects substrate: substrate can adopt amorphous material, macromolecular material, metal oxide ceramic material or semiconductor material;
B prepares metallic film: adopt vacuum deposition method, direct current/radio frequency plasma magnetron sputtering method, or electron beam vacuum deposition method, laser melt physical deposition methods such as method, ion beam sputtering, molecular beam epitaxial growth method, or the method for chemogenic deposit such as metal-organic chemical vapor deposition equipment method, sol-gel process, metal-organic decomposition method, on above-mentioned substrate, prepare metallic film;
The c forming technology: after adopting above-mentioned coating technique to prepare various metallic film samples, with the even glue in the SIC (semiconductor integrated circuit) manufacturing technology, photoetching, etching technics and mask manufacturing process metallic film make electrode, the contact conductor that is connected with each electrode and the electrode external terminal that is connected with each contact conductor;
D absorption biological immune antibody: utilize the bioactivation technology that the electrode surface that adopts above-mentioned forming technology and make is carried out activation processing, absorption one deck activating substance on electrode, then, absorption biological immune antibody on this layer activating substance;
E makes vessel:, employing mark line location face-to-face one by one each electrode that is adsorbed with biological immune antibody on the substrate (2,2 ') that adopts method for preparing placed and is encapsulated in the vessel that a three side sealing closes, the U font wall of recording quantity constitutes, and the external measurement terminal of electrode is positioned at the outside of vessel, and the distance between two substrates is from 0.02 millimeter---10 millimeters.
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|---|---|---|---|---|
| CN101842493B (en) * | 2007-10-31 | 2013-06-19 | 霍夫曼-拉罗奇有限公司 | Electrical patterns for biosensor and method of making |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101842493B (en) * | 2007-10-31 | 2013-06-19 | 霍夫曼-拉罗奇有限公司 | Electrical patterns for biosensor and method of making |
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